cdc25b小分子抑制剂的发现及评价分析化学专业论文.docx

Discovery Discovery and characterization of novel CDC25B inhibitors Cell division cycle 25(CDC25)phosphatases regulate key transitions between cell cycle phases during normal cell division by dephosphorylating and activating the CDK(cyclin-dependent kinase)complexes．And in the event of DNA damage they are the targets of the checkpoint machinery that ensures genetic stability．Since then，CDC25 was reported to be overexpressed in primary tissue samples from various human cancers．Some evidences suggested that the overexpression of CDC25 may contribute to Cancel And down—regulation of CDC25 caused cell cycle arrest and inhibition of cell proliferation．These in turn make them ideal targets for a new anticancer therapy． We purified recombinant CDC25B expressed in Escherichia coli． High—through screening(HTS)assay was set up to find out novel inhibitors of CDC25B from 48000 pure chemicals collected from different sources with wide structural diversity of National Center for Drug Screening．LGH00031 and LGH00045 were discovered and characterized．LGH00031 Was thought to be an irreversible inhibitor,and LGH00045 was a reversible and mixed-type inhibitor of CDC25 with relative good selection．The studies in cellular levels showed that both of the inhibitors caused a G2／M phase arrest which in turn resulted in the inhibition of cell proliferaton,and inhibited dephosphorylation of Tyrl5 of CDKl． The mechanism of the inhibition of LGH00031 against CDC25B was also studied here．The result showed that in vitro the inhibition Was caused by ROS which was generated by the reaction of LGH00031 and DIq．In cultured cells，LGH00031 increased the generation of ROS，and NAC,a precursor of glutathione，impaired the action of LGH00031 in cells．The generation of ROS by the reaction of LGH0003 1 and dihydrolipoic acid Was conjectured，which may be the reason that LGH0003 1 inhibited CDC25B in the absence of DTr in cultured cells． The discovery and characterization of the novel potent and selective inh